Novel 16beta-alkyl steroids and processes



United States Patent 3,116,303 NOVEL lfip-ALKYL STERUHDS AND PROCESfiES David Tauh, Metuehen, Norman L. Wentller, Summit, and Harry L. filates, Madison, Nah, assignors to Merck 8; (30., inc, Railway, N.J., a corporation of New Jersey N0 Drawing. Original application Mar. 19, 1958, Ser. No. 722,390. Divided and this application Mar. 1, 1960, Ser. No. 11,931

2 Claims. (Cl. 260-39145) This invention is concerned generally with novel steroid compounds and with processes of preparing the same, more particularly, it relates to lSfl-alkyl-ll-oxygenated steroids of the pregnane series unsaturated in ring A. it relates also to novel processes for producing 16t3-alkyl steroids as well as la-alkyl steroids.

This application is a division of application Serial No. 722,390, filed March 19, 1958.

The new products of our invention may be represented by the following structural formula:

('30 WW j @j/ A X where 'R is alkyl, R is hydrogen or acyl, R" is hydroxy or oxygen, and X is hydrogen or halogen. The broken line between carbon atoms 1 and 2 indicates that a double bond may be present in this position.

The 16B-alkyl steroids produced in accordance with the present invention possess extremely high anti-inflammatory activity, considerably greater than that of the parent steroids, and are especially effective for the treatment of arthritis and related diseases since they can be administered for their cortisone-like action in extremely low dosage thereby minimizing undesired side eifects. The activity of 16 beta-alkyl steroids is especially novel as all previous group substitution modifications of cortical steroids which have resulted in increased anti-inllam matory activity have involved intnoduction of alpha-substituents.

In preparing our novel chemical compounds, the starting material utilized may bea 3-substituted-1l-oxygenated-l6-pregnene-20-one or 3-substitnted-1l-oxygenated- 16-allopregnene-20-onewhich may be identified by the following structural formula oln3 o=o I ii wnerein X is hydrogen or halogen, Y is alpha or beta hydrogen, Z is oxygen, dioxolane, alpha or beta acyloxy or hydroxy, and R is hydroxy or ox gen as above, with or without a double bond at the (6)-position.

in a preferred embodiment of our invention, 3OL-3C6- ice toXy-16-pregnene-11,20-dione which is represented by the following for-mula is utilized as the starting material. However, it is clear toy wherein R is hydrogen or alkyl.

Upon heating 34x acetoxy-l6a,17a-alkyleneazopregnane-ll,20-dione, there is formed primarily 3a-acetoxy- 16-alkyl-l6-pregnene-1l,20dione which may be represented by the following formula wherein R' is hydrogen or alkyl and exocyclic olefin,

a 3-acetoxy-16-alkylene-pregnane-l1,20-dione, which may be represented by the formulawhere R is hydrogen or alkyl as above.

Treatment of the above isomeric form, namely the 3-acetoXy-16-alkylenepregnane-11,20-dione with an alkali hydroxide yields 16a-a1ky1-3u-hydroxy-16-pregnene- 11,20-dione which has the formulan? log wherein R is alkyl.

Oxidation of 3a-acetoxyl6-alkyl-16-pregnene-11,20-dione, the major isomer obtained above, with an oxidizing agent such as hydrogen peroxide in the presence of sodium hydroxide or peracids such as peroxytrifluoracetic acid results in the formation of 16fl-alkyl-16a,17a-epoxy-3a-hydroxypregnane-l1,2G-dione which has the structural formulard soy wherein R is as above.

Upon treatment of 16fi-alkyl-16a,171x-epoxy-3a-hydroxypregnane-l1,20-dione with a strong acid such as perchloric acid there is formed an olefin mixture of 16- alkyl-3u-17m-dihydroxy-15-pregnene-11,20-dione and 16- alkylene 3a,17u-dihydroxy-pregnane-11,20-dione which may be represented as followsand oj ljignn'" soy wherein R is hydrogen or alkyl and R is as above.

Treatment of the mixture of l6-alkyl-3m,l7a-dihydroxy-lS-pregnene-l1,20-dione and 16-ail:ylene-3a,17adihydroxypregnane-l1,20-dione successively with lithium aluminum hydride and sodium metaperiodate results in a mixture of the endocyciic 16-alky1-3u,11,B-dihydroxy- 15-etiocholene-17-one and the exocyclic, 16-a1kylene-3a, 11fi-dihydroxyetiocholane-17-one having the structural formulawherein R' is hydrogen or alkyl and R is alkyl as above.

Upon treatment of 16fi-alkyl-16a,l7a-epoxy-3tx-hydroxy pregnane-11,20-dione with hydrogen bromide in acetic acid there is formed 3u-acetoxy-16-alkylene-17uhydroxy-pregnane-l1,20-dione, uncontaminated with the 15-pregnene isomer. There is also produced Eta-acetoxy- 16-alkyl-15-bro1no-16-pregnene-11,2O-dionc which may be represented by the following for1nula wherein R is alkyl. The 3a-acetoxy-16-alkyl-15-bromo- 16-pregnene-11,20-dione thus produced is an important intermediate in the preparation of other active steroids.

The 3a-acetoxy-i 6-a1ky1-1S-bromo lG-pregnene-l 1,20- dione may be reacted with pyridine to form 3zx-acetoxy- 16-aikyl-14,16-pregnadiene-11,20'di0ne which has the forrnuia wherein R is alkyl and R is acetyl; the latter compound may be hydrolyzed to the corresponding 3a-o1 which hasthe above formula wherein R is alkyl and R is H.

Hydrogenation of the mixture of 16-alky1-3a,17a-dihydroxy-lS-pregnene-l1,20-dione and 16-alkylene-3a,17adihydroxy-pregnane-11,20-dione with hydrogen in the presence of a hydrogenation catalyst such as palladium on calcium carbonate results in the formation of a mixture of l6oL-21lkYl-30t,17oL-(lihYdlOXYPf63il2ll6-i1,20%0116 and 16s aikyl-3a,l7a-dihydroxypreguane-l1,20-dione w; eh may be represented as follows --0H oz l w R H0 and -0H i j IIO wherein R is as above.

Alternatively, hydrogenation of l6,8-a1kyl-l6u,17aepoxy-3whydroxypregnaue-lLZG-dione under acidic conditions produces directly the above mixture of lfia-alkyl- 3a,17a-dihydroxypregnane-1 1,20-dione and l6 8-aliryl-3 a, 17 a-dihydroxy-pregnane-l 1,20-dione.

Similarly hydrogenation of the pure 16-alkylene-3 a,l7oc dihydroxypregnane-l l,2iCr-dione resulted in the mixture of 160: alkyl 3a,l7a-dihydroxypregnane-l1,20-dior1e and 16fi-alky1-3 65,17 u-dihydroXypregnane-l1,29-di0ne.

At this stage, the mixture may be separated into its components by chromatography or fractional crystallization or other means. As hereinafter described the mixture can be processed in the same manner as either of the two components.

Bromination of the above epirneric mixture or" 16w all yl-3a,l7a-dihydroxypregnane-ll,20-di0ne and 16,8-alkyl-3a,1701-21ihydroxypregnane-l1,2O-dionc results in the formation of an epimerie mixture of l6a-alkyl-2l-brorno- 3or,l7a-dihydroxypregnane-l1,2G-dione and 16 -all\:yl-2lbionic-3a,l7rx-dihydroxypregnane-11,20-dione which may be represented as follows- T O MQU and wherein R is as above.

The above epimeric mixture of ISa-alkyl-Zl-bromo-Su, l7cr-dihydroxypregnane 11,20 dione and 16,8-alky-l-21- 6 bronze-3a,l7aadihydroxypregnane-l1,20-dione may be reaoted with potassium acetate to form an epimeric mixture of l6e-alkyl 3a,l7a,2l trihydroxypregnane-ILZO- dione 21-aeetate and l6fi-aikyl-3u,17a,2l-trihydroxypregmane-ZLZO-dione 21 acetarte which may be represented by the fol-lowing formulae- Clll'IzOAc i= m may and wherein R is as above.

The mixture of 16a-alkyl-17a,21-dihydroxypregnane- 3,1 1,2(Hrione ZI-acetate and 1=6;3-alkyl-17a,Z1-dihydroxypregniane3,l1,20-trione 21-acetate can be separated by chromato gnaphic procedures.

The 16fialkyl-l7a,2l-dihydroxypregrrane 3,11,20 trione Zl-acetate is reacted with bromine to form 16,8-alkyl- 4 hrorno-l7a,2 l-dihydroxypregnane 3,11,20 trione 21- acetate which has the following structural formula- CHzOAC wherein R is alkyl.

The 16p-alkyl 4 br0rno-17a,21-dihydroxypregnane-3, 11,20-trione ZI-acetate obtained above is then reacted with a slurry of sernioarbazide hydrochloride and sodium bicarbonate to form the 3-sernicarbazone of 16,8-alkyll7a,21-dihydroxy-4-pregnene 3,11,20 trione ZI-acetate which may be represented by the following structural formula- CHzOAC n? may wherein R is as above.

The above 3-semioarbazone of 16,8-alky1-17oe21-d ihydroxy-4-pregnene-3,l1,20-trione ZI-acetate is then reacted with, for example, pyruvic acid in aqueous acetic acid to form 16,8-ailkyl-l70,2l-dihydroXy-4-pregnene-3, 11,20-trione ZI-acetate having the following structural formula-- CHrOAc are ozl

wherein R is alkyl as above.

8 Reduction of the 3,20-disernioarbazone of l6fi-a1kyl- 17a,12-dihydroxy-4-pregnene-3,11,20atrione with sodium horonhydride produces the 3,20-disemi-carbaz0ne of 1613- talkyl-llfi,l7a,2l-trihydroxy 4 pregnene 3,20 dione, which has the following formula:

NH2GONHN= I wherein R is as above.

The 3,20-disernicarbazone of 16B-alkyl-llfl,l7a,21-tri-- hydroxyl-pregnene-3,ZO-dione is reacted with pyruvic acid in aqueous acetic acid to remove the semicarbazone groups at the 3 and 20-positions to form 16,8 alky1-11fi, 17a,21 trihydroKy-4-pregnene-3,20-dione which may be chemically represented as follows:

OH2OH wherein R is above. The above compound which may be called 1 65-alky1 hydrocortisone has been found to exhibit marked and effective activity in the treatment of arthritis.

Acetylation of 16 3zalkyl-11fi,l7u,21-tri hydrox-4-pregnone-3,20-dione at the 2l-position is accomplished by reacting this latter compound with acetic anyhdride in the presence of pyridine to form 16,8-a1kyl-11B,17a,21-trihydroxy-4-pregnene-3,ZO-dione Ill-acetate which may be represented as follows:

wherein R is as above.

Other Zl-acylatcs may be prepared by reacting the 21-alcohol with the appropriate acyl anhydrides.

By reacting 16,6-alkyl-11B,17a,21-trihydroXy-4-pregnene-3,20 dione 21-acetate with rnethanesulfonyl chloride in dimethylform-amide there is formed 16fl-alkyl-17a,21- dihydr0Xy-4,9 1 1 -pregnadiene-3,ZO-dione 21-acetate of the following structural formula CIIzOAc wherein R is as above.

Reacting the latter compound with N-bromo-succin- 9 irnide forms 16,8 alkyl-9a-bromo-11fl,17o,21-trihydroxy- 4-nregnene-3,20-dione 2l-acetate represented by the ion mula- CHzOAc HOW/\G E wherein R is as above.

Reacting the latter compound with sodium rnethoxide produces 16,6 alkyl 9,8 11,8 epoxy 1712,21 d-ihydroxy-4-pregnene-3,ZO-dione having the formula--- CIIzOH CHzOAo "OH l j l wherein R is as above. The 21-acetate can be readily converted to the corresponding 21-01, namely, lfl-allryl- 9a fluoro 11 B,17a,2l trihydroxy 4 pregnene 3, 20- d-i-one by hydrolysis. 'ilhis latter compound may also be called 16B-alkyl-9a-iluoroshydrocofiisone and it has been found that this compound is very effective in the treatment of arthritis.

In accordance with this invention, 165-all yl-17a,21- dihydroxy 11 oxygenated 1,4 pregnadiene 3,20- dione and 16 8-alkyl-9a-fluoro-17a,2l-dihydroxy-l bonygenat'ed-1,4-pregnadtiene-3,ZO-dione and the corresponding Zl-acylates can the prepared by reacting respectively 161% alkyl 1711,21 dihydroxy 11 oxygenated pregnane- 3,20-dione 21-acetate and 16fi alkyl-9wfiuoro-17a,21-dihydrox -11-o=;;ygenated pregnane-3,26-dione ill-acetate with bromine to form the corresponding 16fl-" ll yl-2,4-dibrorno 17t,21 dihydroxy 11 oxygenated pregnanc- 3,2( -dione Zi acetate which has the following structural formula:

wherein R is as above and X is hydrogen or fluorine and R is hydroxy or oxygen.

Treatment of these species with dirnethyl aniline in dimethylforrniamide results in the desired 16fl4alkyl-17 11,21- diliydroxy 11 oxygenated 1,4- pregnadiene 3,20- dione ZI-acetate and 16B-alkyl-9a-fluor0-17a,2l-dihydroxy 11 oxygenated 1,4 pregnadiene 3,20 dione 21-acetate which may be represented gnaphioally as followswherein R, R and X are as above. The species represented by the above formula are highly potent anti-inflammatory agents.

The 21-acetate esters of these compounds can be hydrolyzed to the corresponding alcohols, ISfl alkyl-lhlldihydroXy-l l-oxygenated-l ,4-pregnadiene-3,20-di0ne and 165 ialkyl 9a lluoro 17,21 dihydroxy 11 onygenated-l,4-pregnadiene-3,20-dione represented by the following structural formula- CHzOH f/b/ O wherein R and R" are as above.

Alternatively, the 1,4-pregnadiene compounds described above may be prepared from the corresponding 4-pregnene-E-ones by dehydrogenation chemically with selenium dioxide or mierobially by means of Bacillus sphaericus.

The following experimental part illustrates in detail some of the compounds which constitute this invention and methods for their production However, this invention is not to be construed as limited thereby in spirit or in scope since it will be apparent to those skilled in the art that many modifications in materials and methods may be made without departing from the invention.

EXAMPLE 1 Preparation of flcc-Acetoxy-l 6a,] 7a-il4etlzyleneazo- Pregnane-11,20-Di0ne In a 500 ml. 3-neck flash equipped with condenser, dropping funnel and nitrogen inlet were placed 20 g. of potassium hydroxide in ml. of water, ml. of methanol and 100 ml. of ether. A solution of 10 g. of N- rnethyl-Nnntrosotosylaniide in 50 ml. of ether was placed in the dropping funnel.

Diazoniethane was generated by warming the generation flask to 4045 C. and cautiously adding the N- rnethyl-N-nitrosotosylamideether from the dropping funnel. Nitrogen was utilized to sweep the diazomethane into a solution of 20 g. of 3ct-acetoxy-l6pregnene-11,2" dione in 100 ml. of tetrahydrofuran and ml. ether. The process was continued until the steroid solution re mained yellow for several hours. The product, Put-acetoxy-l6a,17a-niethyleneazo-pregnane-11,20 dione largely precipitated from the reaction mixture. After 16 hours, the mixture was filtered, Washed with ether and dried in air. Yield 14.24 g., MP. 186190 C. (dec.), 2nd crop (on concentration) 4.45 g., MP. 187 0., 3rd crop,

11. 1.32 g., MP. 180-190" C. Total usuable material 20.01 g. 90%). LR. A359 5.78 sh, 532,639, xgggf 5.73, 5.82, 6.39.1

Analysis.Calculated for C24H3404N2i C, II, 8.27. Found: C, 69.37; H, 8.01.

EXAMPLE 2 Preparation f 3ot-Acet0xy-1 6-Methyl-16-Pregnene-1] ,2 0- Dione; 3aAcet0xy-16oc,l 7e-Methylenepregnane-l 1 ,2 0- Diane; 3a-Acetoxy-16-Methylenepregnane-11,20-Di0ne and 3 a-Hydroxy -1 6-M ethyl 6 -Pregnene-1 1 ,2 O-Dione 37.4 g. of 3e-acetoxy-l6a,17a-methyleneazopregnane- 11,20-dione was placed in a 500 ml. round-bottom flask and heated by an oil bath in vacuo (pressure 0.6 mm.). A manometer and 12-liter surge flask were in the line between the reaction flask and pump trap. When the bath temperature reached 180 C. the 3a-acetoxy-16a,17amethyleneazo-pregnane-i1,20-dione began to melt with evolution of nitrogen. The maximum pressure reached was 83 mm. After minutes at 180-182" C. the melt was cooled. It had igg 24.9, 13% 191 and was taken up in about 150 ml. of acetone, filtered through diatomaceous earth, concentrated to about 100 ml. and ether slowly added to the boiling solution until crystallization occurred. These crystals of 30t-Z1C6tOXy- 16-methy1-16-pregnene-11,20- dione weighed 19.0 g., MP. 165-168 C.;

xg jg 249, 2% 234 2nd crop 5.3 g., M.P. 160-164" 0,

3rd crop 5.0 g., M.P. 142-153" C.,

xCHaOH 249, 13% 156 max.

Total yield of material with E% 210 was 70%. The analytical sample had M.P. 165-167" C.

EXAMPLE 3 Preparation 0 f 1612,] 7a-Epoxy -3 a-Hydr0xy 65- Me thy ipregnane-l 1 ,2 0-D ione A solution of 20.0 g. of 3a-acetoxy-16-rnethyl-16-pregnene-l1,20-dione dissolved in 600 ml. of methanol, was cooled to 18 C., and 80 ml. of 30% hydrogen peroxide followed by 80 ml. of 2.5 N sodium hydroxide were added. Considerable material precipitated from solution, but all redissolved on stirring the reaction mixture at C. for minutes. The solution was kept at 15 -20 C. for 18 hours at which time the ultra-violet maximum at 249 had completely disappeared. Then 600 ml. of saturated salt water was slowly added, the crystalline precipitate was filtered, washed with water, and dried in air and in vacuum. The 166:,17a-epoxy-3ot-hydroxyl6l3-methylpregnane-11,20-di011e thus formed weighed 12 17.36 g. (93%); MP. 176-177" C., hexagonal prisms, MP. 178-180" C., from acetone-ether.

AnaZysis.-Calculated for C H O C, 73.30; H, 8.95. Found: C, 73.23; H, 9.12.

Room temperature acetylation gave the 3a-acetoxy- 16oz,17a-epoxy-16fl-methylpregnane-11,20-dione having a melting point of 162-164" C.

Analysis.-Calculated for C H O C, 71.65; H, 8.51. Found: C, 71.65; H, 8.25.

EXAMPLE 4 Preparation of an Olefin Mixture of 3a,]7ot-Dilzydr0xy- 16-Melhyl 15 Pregnene-ZLZO-Dione and 306,17Ct-Dilzydroxy-l 6-Methylenepregnane-l 1,20-Di0ne To a solution of 2.69 g. of l6a,l7a-epoxy-3a-hydroxyl6/3-methyl-pregnane-11,20-dione in 55 ml. dioxane was added 27 ml. of 2 M aqueous perchloric acid. The clear solution was kept at 25-30" C. for 65 hours. Cold water, (175 ml.) was added, the slurry chilled to 8 C. and filtered after 30 minutes. The precipitate, containing a mixture of 312,171! dihydroxy 16-methyl-15-pregnene- 11,20-dione and 31x,17u-dihydroxy-lfi-methylene-pregnane was washed with water, and dried in air and finally at 50 C. in vacuum. Yield: 2.27 g. (94.5%), MP. sintering at about C., melting at 158-167" C. The relative proportion of 3(1,17u-dihydroxy-16-methyl-15-pregnene- 11,20-dione and 30,1706 dihydroxy-lG-methylenepregnane-11,20-dione is estimated to be of the order of 1:1.

EXAMPLE 5 Preparation of 3a,] Ifi-Dihydroxy-I6-Methyl-15-Eti0cholane-1 7-0ne and 3a,]1,B-Dihydroxy-I6-Methylene-Eti0- cholane-Z 7-One The conjugate C=O peaks at 5.80 and 5.87 were of equal intensity indicating the proportions to be 1:1.

Pore 3a,11 8 dihydroxy 16 methylene-etiocholane-17- one max.

max.

011013 max.

was prepared similarly from pure 3a-acetoxy-16-methylene-lh-hydroxy-pregnane-11,20-dione obtained by hydrogen bromide opening of the oxide, 3a-acetoxy-16a, 17aepoxy-16fi-methyl-pregnane-11,20-dione (see Example 6).

EXAMPLE 6 Preparation of 3a. Acetoxy-Z7a-Hydr0xy-16-Methylene- Pregnane-11,20-Di0ne and 31x Acet0xy-15-Br0m0-16- M ethyl-.1 fi-Pregnene-l 1,20-Di0ne To a stirred solution of 5.05 g. of 3a-E1C6tOXy-16a,l7ot epoxy-16B-methyl-pregr1ane-11,20-dione in 75 ml. of acetic acid maintained at 10-15 C. was added 25 ml. of cold 15% hydrogen bromide in acetic acid. After 35 minutes at 10-15 C. the mixture was concentrated to dryness in vacuo (temperature 15 C.) and the residue chromatographed on 200 g. of neutral alumina. From the 60:40 petroleum ether-benzene eluates there was obtained 2.10 g. of 30 acetoxy-l5-bromo-16-methyl-16-pregueneis 11,20'dione; rectangular prisms from ether-acetone, MP. 130-135 C., dec.,

[04199 +13 0:101 ig gg 250 my, E=9,000;

[01] g 5.2 (C. 1.00); rigid 2.95, 5.75, 5.82, 5.87

Analysis-Calculated for C I-1 0 C, 71.61; H, 8.51. Found: C, 71.51; H, 8.07.

EXAMPLE 7 Preparation of 3a-Acet0xy-16-Methyl-14J 6- Pregnadiene-l 1 ,ZO-D ione 42 mg. of 35i-acetoxy-15-bromo-16-methy1-16-pregnene- 11,20-dione was dissolved in 2 ml. of pyridine. After 18 hours at 25 C. the mixture was concentrated to dryness in vacuo. The solid residue of 3a-acetox -16-methyl- 14,16 pregnadiene 11,20 dione was crystallized from ether; prismatic needles, MP. 180-185 C;

XCHBOH 304 m E: 11,000

max.

EXAMPLE 8 A solution of 3.05 g. (8.47 millimols) of the olefin mixture of 31:4,1701 dihydroxy '16 methyl-15-pregnene- 11,20- dione and 3a,l7a-dihydroxy 16 7 methylene-pregnane-lLZG-dione in 80 ml. of methanol was reduced in hydrogen at 1 atmosphere and 25 C. in the presence of 2.0 g. of 25% palladium-calcium carbonate catalyst. Modification of the hydrogenation conditions, pH, solvent, catalyst, etc. alters the isomer ratio significantly. Uptake of the calculated amount of hydrogen was complete in 45 minutes. The mixture was stirred an additional 30 minutes and filtered through diatomaceous earth. The colorless filtrate was taken to dryness and crystallized from ether: a mixture of 3a,17a-dihydroxy-16e-methylpregnane-dLm-dione and 3a,17u-dihydroxy-loo-methylpregnane-l1,20-dione was obtained and weighed 3.05 g., "tutoring at 150 C., melting at 166182 C.

product consists of 311,17ix-dil1ydroxy-16a-methylpregnane-llittdione and 3a,l7c-dihy droxy-t1GS-methylpregnone-11,20-dione in the ratio oa. 7:3 as determined by the amounts of end product isolated below.

EXAMPLE 9 Alternate Preparation of 311,17a-Dihydr0xy-16a-Methylpregnane 11,20 Dione and 3a,]7ci Dihydroxy-Mfi- M ethyl pregnane-l 1 ,ZO-Dione A solution of 10.0 g. of 16c,17a-epoxy-3a-hydroxy- 16,8-hethylpregnane-11,ZO-dione in 180 ml. of dioxanc and 112 ml. of 2 M aqueous perchloric acid was prepared. The solution was hydrogenated at 25 C. and 1 atmosphere pressure in the presence of 4.4 g. of 5% palladium on charcoal catalyst. Uptake of one mole of hydrogen was complete in 10 hours. The catalyst was removed by filtration and 300 ml. of 70% saturated sodium chloride was added to the filtrate. After being kept at 0 C. for 30 minutes, the precipitated mixture of 3a,l7a dihydroxy 16a methylpregnaned1,20-dione and 3ci, 17a-dihydroxy 16,8 methylpregnane-l1,20-dione was filtered, washed with water and dried in air. Yield: 9.26 g. Solubility analysis in benzene indicated the prod- 14 not to be 75% 3a,17a-dihydroxy-1Gwmethylpregnane- 11,20-1dione and 25% 3e,17a-dihyd-roxy-16/3-methylpregn ane-11,20-dione.

EXAMPLE 10 Separation of 311,177; Dihydroxy-Ma Methylpregntme- 11,20-Dione and 3a,17a-Dihydr0xy 16,8 Methylpregnane-lI,20-Di0ne One gram of the above hydrogenation product, 3a,17adihydroxy l6a-methylpregnane-11,20-dione and I3a,17a-di hydroxy-lfifl-methylpregnanedl,20 dione, was chromatographed on g. of activated magnesium silicate. The 100% chloroform elu ateis gave 3a,17a-dihydroxy-16ame-thy1pregnane-11,20- dione, MP. 188-191 C., identical with authentic material by mixed melting point and infrared spectral comparisons. The 5% methanol-chloroform eluates gave 3a,17a-dihydroxy-16B-methylpregnane- 11,20di0ne hexagonal plates from benzene-ethylacetiate, M.P. 192197 C.;

Ai1a lysis.Ca1culated for C H O C, 72.89; H, 9.45. Found: C, 72.97; H, 9.25.

The la-methyl and 16/3-methylpregnar1es respectively, ha d R values of 0.55 and 0.35 when chromatographed on Whatman #1 filter paper in the benzene-formamide systern.

As described hereinafter, 3a,17a-dihydroxy-16,8-methylpregnane-11,20-dione was brominated at position 21 in exactly the same way as the mixture of 16a and 16emethyl compounds to give 21-bromo-3a,17 z-dihydroxy- -methylpregnane-11,20-dione. This latter compound was similarly converted to 3a,17oi,21-trihydroxy-16fi-methylpregnane-11,20-dione 21-acetate. This compound was then similarly oxidized at position 3 to give 17a,2l-dihydroxy-165-methyl-3,11,20-trione 2lacetate.

EXAMPLE 11 Preparation of a Mixture of 3 ai-A cetOxy-J 7a-Hydroxy- 16a-MethyI-Pregnane-1 1,20-Di0ne and 3a-Acet0xy-1 70c- Hydroxy-J Gcthyipregnmze-Z 1 ,ZO-D i 0 us To a mixture of 3m,17a-dil1ydroxy-16a-rnethylpregnane-11,20-dione and 3e,17a-dihydroxy-163-rnethylpregnane-11,20-d-ione dissolved in pyridine was added acetic anhydride, at 20 C. A mixture of 30i-acetoxy-17a-hydroxy-1Sa-methylpregnane-l1,20-dione and 3e-acetoxy- 17o: hyd'ro xy-1fi-methylpregnanel1,20-dione separated from solution and was recovered. These prisms sintered at C. and melted at 179 C.

EXAMPLE 12 Preparation of a Mixture of 21-.3r0mo-3a,17a-Dihydroxyloa-Methylpregnane 11,20 Dione and 2l-Bromo-3ix, 17a-Dir'iydr0xy-1 6 5-114 etitylpregrmne-l 1 ,ZO-Dione A solution of 3.50 g. (9.7 millirnols) of a mixture 301,17 di hydroxy 16c: methyipregnaue 11,20 dione and 3a,17 x dihydro:-;y-16-methylpregnane-11,20-dionc obtained as above in 40 ml. of chloroform was warmed to 40-45 C. A solution of 1.76 g. (11 miiiimois) of bromine in 25 ml. of chloroform Was added drcpwise to the stirred solution such that the color was not darker than pa le yeilow (ca. 2 drops/see, total time, 1 hour). The nearly coloriess solution was cooled to 20 C. and 200 ml. of ether was added. The mixture was extracted with excess cold 5% potassium. bicarbonate solution, sodium b isu ifite solution, and Water and dried over 11 agnesium sulfate. The colorless residue after removal of solvent, a mixture of 21-bromo-3a,l7cx-dihydroxy-16amethylpregnam-i-LZO-dione and 21-brorno-3 a,17ocdil1ydroxy-lfi-methyipregnane-l1,20-dione (4.30 g.) gave a positive tetrazoiium test. Paper chromatography (benzene-formamide) resolved material into 2 spots.

17 maintained at 45 C. (for 17 hours. It was then cooled to 20 C. and 50 ml. of 50% saturated aqueous sodium chloride was added. After 2 hours at C. the precipitate of the 3,20-disemicarbazidone of 17a,21-dihyd-roxy- 16/3-methyl 4 pregnene-3,11,20-trione 21-acetate was filtered, washed with water until free of chloride ion and dried in air. Yield over 90%.

EXAMPLE 19 Preparation of the 3,20-Disemicarbaz0ne of 11B,17a,21-

Trihydroxy-l ofi-Methyl-4-Pregnene-3,20-Dione To a stirred solution of 600 mg. of the 3,20-disemicarbazone of 17a,21-dihydroxy-16B-methyl-4-pregnene-3, 11,20-trione 21-acetate in 30 m1. of tetrahydrofuran and 11 ml. of water under nitrogen was added 200 mg. powdered sodium borohydride. The stirred suspension was refluxed 45 minutes and then cooled to 15 C. Aqueous acetic acid (3 ml. of 30%) was added cautiously and most of the tetrahydrofuran was removed in vacuum. Addition of 5 ml. of methanol and 5 ml. of water induced the product to crystallize. Following addition of ml. of a saturated sodium chloride solution and aging at 0 C. the product 3,20-disemicarbazone or 11B,17a,21-trihydroxy-16/i-methyl 4 pregnene 3,20 dione was filtered, washed with water, and dried in air.

EXAMPLE 20 Preparation of 11fl,17a,21-Trihydr0xy-l 6 B-M ethyl-4- Pregnene-3,20-Dione To a solution of 510 mg. of reduced 3,20-disemicarbazone of 11B,l7a,21 trihydroxy-16B-metl'1yl-4-pregnene-3, 20-dione in 5 ml. of acetic acid was added 1.20 ml. of water and 0.50 ml. of pyruvic acid. The solution was kept at 25 C. for eighteen hours. Water (20 ml.) was added, and the mixture was extracted thoroughly with chloroform. The chloroform extract was dried over magnesium sulfate and taken to dryness. The residue was crystallized from acetone-ether to give pure llfi,l7a,21- trihydroxy-16,8methyl-4-pre@ene-3,20-di0ne.

EXAMPLE 21 Preparation of 115,] 7a,21-Trihydroxy-16B-Methyl-4- Pregnene-3,20-Dione 21 -Acetate The product of the previous Example 20 was acetylated at C21 as follows: A solution of 100 mg. of 1lfi,17oc,21- trihydroxy-l6fi-methyl-4apregnene-3,ZO-dione in 100 ml. of pyridine and 0.5 ml. of acetic anhydride was prepared. After 18 hours at 25 C., the solution was taken to dryness in vacuo and the solid residue purified by crystallization in acetone-ether to give 11,8,17a,21-trihydroxy- 16,8-methyl-4pregnene-3,20-dione 21-acetate.

EXAMPLE 22 Preparation of 1 7a, 21-Dil2ydroxy-16,8-Metlzyl-4,9(1 1)- Pregnadiene-3,20-Di0ne 21 -Acetate A solution of 400 mg. of l1B,17a,21-trihydroxy-16fimethyl-4-pregnene-3,20-dione 21-acetate in 2.0 ml. dimethyl formamide, 0.8 ml. of pyridine and 0.4 ml. of methanesulfonyl chloride was kept at 75 C. for one hour. The mixture was cooled, water added, and the precipitate filtered, washed "with water and dried in air. The desired l7a,21-dihydroxy-16,8-methyl-4,9(11)-pregnadiene-3,20-dione 21-acetate was purified by chromatog raphy on alumina (20 g.) and elution of the column with benzene. Crystallization of material eluted by benzene gave pure 17a,21-d=ihydroxy-16,8-methyl-4,9(11)-pregnadiene-3.20-dione 21-acetate.

EXAMPLE 23 Preparation of 9a-Br0mo-1 1 [3,1 7a,21-Trihydr0xy-16,8-

M ethyl-4-Pregnene-3,20-Di0ne 21-Acetate To a mixture of 620 mg. of 17a,21-dihydroxy-16,8- methyl 4,9 1 1) pregnadiene 3,20-dime 21-acetate and 330 mg. of N-b-romosuccinimide in 10 ml. of dioxane and 3.2 ml. of water cooled to 10 C. was added 1.8 ml. of cold 1 M aqueous perchloric acid. The mixture was stirred at 15 C. for 3 hours. Excess N-bromosuccinimide was destroyed by addition of aqueous sodium thiosulfate and most of the dioxane was removed in vacuo. About 30 ml. of water was added and crystalline bromohydrin, 9a=bro rno 1lfl,l7o,21 trihydroxy-16p-methyl-4- pregnene-3,20-dione ZI-acetate, was filtered, washed with water, and dried in air.

EXAMPLE 24 Preparation of 95,] 1 ,B-Epoxy-l 7a,2I-Dihydr0xy-16fl- M ethyl-4 -Pregnene-3 ,ZO-Di one 21 -A cetate To a stirred solution of 100 mg. of the 9a-bromo-11B, 17a,21-trihydroxy-165-methyl-4-pregnene-3,20-dione 21- acetate in 3 ml. of tetrahydrofuran and 1 ml. of methanol under nitrogen was added 1.02 ml. of 0.215 N methanolic sodium methoxide. After 10 minutes at 25 C. 0.2 ml. of acetic acid was added and the methanol removed in vacuo. The residue was acetylated with 1.00 ml. of pyridine and 0.5 ml. of acetic anhydride at 60 C. for 70 minutes. The mixture was taken to dryness in vacuo, water added, and the product extracted into chloroform. The residue was crystallized from etheracetone to give pure 9,8,1lB-epoxy-17a,2l-dihydroxy- 165-methyl-4-pregnene-3,20-dione 2l-acetate.

EXAMPLE 25 Preparation of 9a-Flnoro-Z 1,8,1 7u,21-Trihydr0xy-16p- M ethyl -4 -Pregnene-3 ,2 O-Dione 21 -A cetate To a solution of 200 mg. of 9fi,1lfi-epoxy-17u,21-dihydroxy-16fi-methyl-4-pregnene-3,20-dione 21-.acetate in 2 ml. of chloroform and 2 ml. of tetrahydrofuran in a polyethylene bottle at -60 C. was added 2 ml. of a 2:1 (by Weight) mixture of anhydrous hydrogen fluoride and tetrahydrofuran. After 4 hours at 10 C. the mixture was cooled to 60- C. and cautiously added to a stirred mixture of 30 ml. of 25% aqueous potassium carbonate and 25 ml. of chloroform kept at 5 C. The aqueous phase was further extracted with chloroform and the latter phase washed with water and dried over magnesium sulfate. The residue on crystallization from acetone-ether gave pure LfiUOI0-11[3,17a, 2l-trihydroxy-l6p-methyl-4-pregnene-3,20-dione 21 acetate.

EXAMPLE 26 Preparation of 9a-FZa0r0-1 1 5,1 711,21 -Trihydr0xy-1 6,8- filethyl-4 Pregnelie-3,20-Dione To a stirred solution of 110 mg. of 9ot-fiuOI'O-11fi3,17oc, 2l-trihydroxy-16,8-methyl-4-pregnene-3,20-dione 21 acetate in 5 ml. of methanol under nitrogen at 25 C. was added 1.00 ml. of 0.26 M methanolic sodium methoxide. After 15 minutes 0.2 ml. of acetic acid in 1 ml. of water was added and the mixture concentrated nearly to dryness. The residue was taken up in ethyl acetate and the ethyl acetate solution was washed with water, dried over magnesium sulfate, and concentrated to dryness. Crystallization of the residue from ethyl acetate gave pure 9a-fluoro-l1B,17a,21-trihydroxy-16 3-methyl-4-pregnene-3,20-dione.

EXAMPLE 27 Preparation of 9a-Clzl0ro-1 1 13,1 7a-21Trihydroxy-16[3- M ethyl-4-Pregnene-3,20-Dione 21 Acetate To a solution of mg. of 9,6,11;8-epoxy-17a,21-dihydroxy-l6fl-methyl-4-pregnene-3,20-dione 21-acetate in 4 ml. of chloroform was added 5 ml. of concentrated hydrochloric acid. The two-phase mixture was stirred at 25 C. for 1 hour. Addition of water and chloroform extraction gave a crude crystalline product which is partly deacetylated. Treatment with 1 m1. of pyridine and 0.5 ml. of acetic anhydride at 25 C. for 18 EXAMPLE 28 Preparation of 2,4-Dibrm0-1 7a,2l Dihydr0xy-16fi- Methylpregnane-3,11,20-Tri0ne 21 -Acetate To a stirred solution of 682 mg. of 17a,21-dihydroxy- 16 8-methylpregnane-3,11,20-trione 21-acetate (prepared as described in Example 17) in 20 ml. of chloroform and 2.25 ml. of acetic acid maintained at 20 C. was added dropwise one-half of a solution of 540 mg. of bromine in 2 ml. of chloroform and 3 ml. of acetic acid. The mixture was warmed to 0 C. and the remainder of the bromine added. Sodium acetate (0.4 g.) in 2 ml. of water was added followed by 20 mg. of sodium sulfite. The mixture was concentrated in vacuo to remove the chloroform and 20 ml. of water was added. The white powdery precipitate of 2,4-dibromo-17a,21-dihydroxy 16,8 methylpregnane-3,l1,20-trione 2l-acetate was filtered, washed with water and dried in air. Yield: 920 mg, MP. 122-130 C. dec.

EXAMPLE 29 Preparation of 1 7a,21-Dihydr0xy-1 6,8-Methyl-1,4- Pregnadiene-3,I1,20-Tri0ne 21-A cetate To a solution under nitrogen of 900 mg. of the 2,4-dibromo l7oc,21 dihydroxy 16,8 methylpregnane 3, 11,20-trione 21-acetate in 5 ml. dimethyl formamide was added 200 mg. of sodium bromide. After 1 hour at 25 C., 1 ml. of dimethylaniline was added and the mixture maintained at 135 C. for 2% hours. The mixture was cooled, added dropwise to dilute hydrochloric acid, and solid, crude product filtered, washed with dilute hydrochloric acid, Water and dried in air. Treatment with charcoal, followed by crystallization from acetone gave 1711,21 dihydroxy 16B methyl 1,4 pregnadiene 3, 11,20-trione 21-acetate, MP. 230-233 C.,

onion maX.

236 my. (E=15,300) xCHOh Analysis.Calculated for C H O C, 69.55; H, 7.30. Found: C, 69.25; H, 7.25.

EXAMPLE 30 Preparation 0 115,1 7a,21-Trihydr0xy-1 6,8-Methyl-1,4 Pregnadiene-3,20-Di0ne 21-Acetate To 100 mg. of 1l/i,17a,21-trihydrotxy-d6B-methyl-4- pregnene-3,20-dione ZI-acetate in 5 ml. of t-butanol and 0.1 ml. of acetic acid was added 50 mg. of selenium dioxide. The mixture was refluxed under nitrogen 18 hours, 50 mg. of selenium dioxide was added and the mixture refluxed an additional 24 hours. The mixture was filtered, and the filtrate taken to dryness. The residue was taken up in ethyl acetate and washed successively with aqueous sodium bicarbonate, ammonium sulfide, dilute ammonia water, water, dilute hydrochloric acid and water and dried over magnesium sulfate. It was then treated with activated charcoal and concentrated to dryness. Crystallization of the residue from acetone-ether gave pure 115,17a,21-trihydroxy-165-methyl-1,4-pregnadiene-3,20-dione 21-acetate.

EXAMPLE 32 Alternately, the 1,2-dehydrogenation of 1lfi,l7a,21-trihydroxy-l6fl-methyl-4-pregnene-3,20-dione 21-acetate can be carried out microbiologically by means of Bacillus sphaericus to yield 11B,17u,21-trihydroxy-16[3methyl- 1,4-pregnadiene-3,20-dione, which on acetylation with acetic anhydride-pyridine gives 11B,17a,21-trihydroxy- 16fl-methyl-1,4-pregnadiene-3,20-dione 21-acetate.

EXAMPLE 33 Preparation of 9a-Flu0r0-1 1,6,1 7a,21-Trihydroxy-16fi- M ethyl-1 ,4-Pregnadiene-3,2 O-Dione 21 -A cetate In a similar manner, 100 mg. of 9u-flu0r0-1 1B,17a,21- trihydroxy-16,8-methyl-4-pregnene-3,20-di0ne 21 acetate was treated with selenium dioxide to produce the corresponding 90c-fi1lO1'0-l l/3,17oa,21 trihydroxy 16B methyl- 1,4 pregnadiene 3,20 dione 21 acetate. Alternately, Bacillus sphaericus may be utilized. The 21-alcohol was prepared from the 21-acetate as in Example 26 or Example 30.

Various changes and modifications may be made in the present invention, certain preferred embodiments of which are herein disclosed, without departing from the scope thereof; to the extent that these changes and modifications are within the scope of the appended claims, they are to be considered a part of this invention.

We claim:

1. The process which comprises reacting a mixture of 16a-alkyl-3oc,17oz,2 l-trihydroxypregnane 11,20-dione 21- acylate and 16,8 alkyl 3a,17a,21 trihydroxypregnane- 11,20-dione 21-acylate with N-bromosuccinimide to form a mixture of 16a-alkyl-17a,21-dihydroxypregnane-3,11,20- trione 21-acylate and 16fl-alkyl-17a,21-dihydroxypregmane-3,11,20-trione 21-acy1ate and chromatographically separating the said latter mixture.

2. The process which comprises reacting 16fl-alkyl- 3a,17a,21 trihydroxypregnane 11,20 dione 2l-acylate With N-bromosuccinimide to form 16/3-alkyl-17a,21-dihydroxypregnane-3,11,20-trione 21-acylate.

References Cited in the file of this patent UNITED STATES PATENTS 2,783,254 Gould et a1 Feb. 26, 1957 2,832,773 Farrar et a1. Apr. 29, 1958 2,838,498 Magerlein et al June 10, 1958 2,954,326 McAleer et al Sept. 27, 1960 FOREIGN PATENTS 712,058 Great Britain July 14, 1954 773,016 Great Britain Apr. 17, 1957 788,306 Great Britain Dec. 23, 1957 OTHER REFERENCES Klyne et al.: J. Chem. Soc. (1956), pp. 4825-28. Wendler et al.: J.A.C.S. 73, 3818-3820 (1951). Meystre et al.: Helv. Chim. Acta 39, 734-742 (1956). Ringold et al.: J. Org. Chem. 21, 239-40 (1956). Szpilfogel et al.: Rec. Trav. Chem. 75, 475- (1956). Marker et al.: J.A.C.S. 64, 1280 (1942). 

1. THE PROCESS WHICH COMPRISES REACTING A MIXTURE OF 16A-ALKYL-3A,17A,21-TRIHYDROXYPREGNANE - 11,20-DIONE 21ACYLATE AND 16B-ALKYL - 3A, 17A,21 - TRIHYDROXYPREGNANE11,20IDIONE 21-ACYLATE WITH N-BROMOSUCCINIMIDE TO FORM A MIXTURE OF 16A-ALKYL-17A,21-DIHYDROXYPREGNANE-3,11,20TRIONE 21-ACYLATE AND 16B-ALKYL-17A,21-DIHYDROXYPREGNANE,3,11,20-TRIONE 21-ACYLATE AND CHROMATOGRAPHICALLY SEPARATING THE SAID LATTER MIXTURE. 